Synthesis of multiple antigenic peptides: strategies and limitations
Dendrimeric platforms such as MAPs can be synthesized by a solid phase method or by conjugation. Dendrimeric platforms such as MAPs can be synthesized either entirely by solid-phase methods (SPPS, direct approach) or by conjugation in a solution of preformed, SPPS-made building blocks (indirect approach).
SPPS is the preferred method by LifeTein. The synthesis approach requires a branched poly-lysine core. Each branch is elongated into the corresponding epitope by stepwise SPPS. The disadvantage of this approach is that synthetic errors could happen and cause microheterogeneity in the final materials. However, the cost is lower and less time-consuming than the indirect approach. For very long linear peptides, it is more advantageous to use the SPPS method.
The MAP synthesis may not always meet with success. The solubility of the peptide epitope can also become an issue and is difficult to predict for long epitopes. It is recommended to carefully design and analyze the linear epitope before MAP synthesis.
Studies showed that synthesis with Ahx linker in the lysine core had a better-isolated yield. It is possible that the flexibilities of Ahx help in keeping peptide chains properly solvated during synthesis, preventing aggregation, and hence increasing the amount of viable growing peptide sequences.
Did you know that 15%–40% of all interactions in the cell are mediated through protein-peptide interactions?
The following peptide designs are powerful strategies to learn about the general features of optimal peptide sequences binding to a given domain or protein.
- Enhanced solubility: Analyze charged residues for better solubility.
- Control peptides: Scrambled peptide as a control.
- CPP peptide: TAT sequence for cell penetration.
- Spacer or Linker: A GG, or PEG linker to increase flexibility.
- D amino acid peptide: Avoid degradation.
- Biotin conjugation: Biotin/avidin of pull-down assay for your target.